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Human α 1-Antitrypsin Gene Transfer to In Vivo Mouse Hepatocytes

1996; Mary Ann Liebert, Inc.; Volume: 7; Issue: 4 Linguagem: Inglês

10.1089/hum.1996.7.4-531

1557-7422

Salvador F. Aliño, María Bobadilla, J. Crespo, M. Lejarreta,

Virus-based gene therapy research

The in vivo gene transfer to mouse hepatocytes of pTG 7101, a plasmid containing the full-length gene encoding human α1-antitrypsin (α1-AT) DNA, has been studied by iv administration of recombinant DNA (100 ng/mouse) encapsulated in large and small liposomes. Our results from immunohistochemical liver sections and cytophotometric analysis of hepatocyte chromophore absorbance indicate that human α1-AT was expressed in liver parenchymal cells from mice treated (48 hr before) with DNA encapsulated in small liposomes, and this effect remained for at least 2 weeks. In contrast, the efficiency was greatly limited when large liposomes were used as a vehicle for gene transfer. Additional experiments were performed to study using an ELISA procedure the presence in mouse plasma of human α1-AT from mice treated with encapsulated plasmid in small liposomes or small empty liposomes plus free DNA. According to the immunohistochemical data, the results indicate that detectable α1-AT can only be observed in plasma from mice treated with encapsulated plasmid in small liposomes. We studied the effect of in vivo gene transfer into mouse hepatocytes of pTG 7101, a plasmid containing the full-length human α1-antitrypsin (α1-AT) gene, by iv administration of recombinant DNA encapsulated in large versus small liposomes. Immunohistochemical and cytophotometric analysis of hepatocytes showed human α1-AT expression only in the liver of mice treated with DNA encapsulated in small liposomes. This expression persisted for 2 weeks. In contrast, when large liposomes were used, the efficiency was greatly reduced. Additionally, plasma levels of human α1-AT could be detected only in mice injected with the plasmid encapsulated in small liposomes.